Siinfekl peptide

Manufactured by Thermo Fisher Scientific

SIINFEKL peptide is a synthetic peptide sequence commonly used in immunological research. It serves as a model antigen for studying T-cell responses and immune system function. The peptide can be utilized in various in vitro and in vivo experimental settings to investigate topics such as antigen presentation, T-cell activation, and immune system dynamics.

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Lab products found in correlation

Rat igg, by BioXCell (1 mentions) Anti cd137 clone 3h3, by BioXCell (1 mentions) C57bl 6 mice, by Jackson ImmunoResearch (1 mentions) Anti cd3 28 beads, by Thermo Fisher Scientific (1 mentions) Carboxyfluorescein succinimidyl ester (cfse), by Thermo Fisher Scientific (1 mentions) Ionomycin, by Thermo Fisher Scientific (1 mentions) Siinfekl, by Merck Group (1 mentions) Cd8a ly 2, by Miltenyi Biotec (1 mentions) Incomplete freund s adjuvant, by Merck Group (1 mentions) Ls column, by Miltenyi Biotec (1 mentions) Quadromacs separator, by Miltenyi Biotec (1 mentions) Cd11c microbeads, by Miltenyi Biotec (1 mentions)

5 protocols using siinfekl peptide

Adoptive Transfer of OT-I T Cells and Costimulation

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C57BL/6 mice were purchased from The Jackson Laboratory (Bar Harbor, ME). OT-I RAG^−/− transgenic mice were bred in-house. All mice were maintained in the UConn Health animal facility in accordance with National Institutes of Health guidelines. Spleen plus lymph node preparations from CD45.1⁺ RAG^−/− OT-I transgenic mice containing 5 × 10⁵ CD8⁺ T cells were adoptively transferred i.v. into CD45.2⁺ C57BL/6 recipients that were subsequently injected i.p. with 50 μg SIINFEKL peptide (Life Technologies; Grand Island, NY) and 30 μg Rat IgG or 20 μg anti-CD134 (clone OX86, Bio X Cell, Lebanon, NH) plus 10 μg of anti-CD137 (clone 3H3, Bio X Cell) agonists the following day as previously described (19 (link)) (20 (link)).

Tsurutani N., Mittal P., St Rose M.C., Ngoi S.M., Svedova J., Treadway F.B., Laubenbacher R., Suárez-Ramírez J.E., Cauley L.S., Adler A.J, & Vella A.T. (2015). Costimulation endows immunotherapeutic CD8 T cells with IL-36-responsiveness during aerobic glycolysis. Journal of immunology (Baltimore, Md. : 1950), 196(1), 124-134.

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Co-Culture of OT-1 T Cells

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Equal numbers of OT-1 (CD45.1) and OT-1 x Rag2^−/− (CD45.2) T cells were co-cultured in complete RPMI + 10% DBS with 1:1000 2-mercaptoethanool (Sigma) containing 1 μM SIINFEKL peptide or anti-CD3/28 beads (Lifetechnologies).

Karo J.M., Schatz D.G, & Sun J.C. (2014). The RAG recombinase dictates functional heterogeneity and cellular fitness in natural killer cells. Cell, 159(1), 94-107.

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Adoptive Transfer of OTI Cells for Tumor-Specific Cytotoxicity

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WT or IFNγRKO OTI cells were transferred in tumor-bearing WT mice between 5 and 7 days after B16-OVA cell engraftment when the tumor was palpable. After 7 days of OTI adoptive transfer, mice were injected with a 1:1 ratio of Ova-targetable and non-targetable cells. Splenocytes from naïve B6 mice were isolated and split into two equal pools. Targetable cells were loaded with SIINFEKL peptide (1 µg/mL) and further labeled with high concentration CFSE (2 µM - Invitrogen). Non-targetable cells were not loaded but labeled with low CFSE (0.2uM). Peptide loading was performed in complete RPMI at 37 °C for 1 h. Cell labeling was done in PBS for 15 min at 37 °C. After cell preparation, cells were washed with PBS and pooled together for intravenous injections. After 24 h spleens were harvested and processed for flow cytometry analysis.

Mazet J.M., Mahale J.N., Tong O., Watson R.A., Lechuga‐Vieco A.V., Pirgova G., Lau V.W., Attar M., Koneva L.A., Sansom S.N., Fairfax B.P, & Gérard A. (2023). IFNγ signaling in cytotoxic T cells restricts anti-tumor responses by inhibiting the maintenance and diversity of intra-tumoral stem-like T cells. Nature Communications, 14, 321.

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Cytokine Production in Immune Cells

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In order to measure the capacity of isolated immune cells to produce cytokines, before staining, cells were incubated at 37 °C for 5 h with 2 μM brefeldin A (BFA) to stop Golgi secretion, 1 μM SIINFEKL peptide, 0.04 μM PMA, and 0.5 μM ionomycin (Invitrogen).

Noffsinger B., Witter A., Sheybani N., Xiao A., Manigat L., Zhong Q., Taori S., Harris T., Bullock T., Price R, & Purow B. (2021). Technical choices significantly alter the adaptive immune response against immunocompetent murine gliomas in a model-dependent manner. Journal of neuro-oncology, 154(2), 145-157.

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Isolation and Activation of Immune Cells for In Vivo Studies

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Isolation of CD8⁺ T cells or CD11c⁺ DCs was performed by immunomagnetic selection from splenocytes using Manual MACS Cell Separation Technology (QuadroMACS Separator, LS columns, CD8a [Ly-2] and CD11c MicroBeads; Miltenyi Biotec), according to the manufacturer’s instructions. For in vitro experiments, SIINFEKL peptide (Invitrogen) was added at a concentration of 5 μM. In competitive in vivo experiments, 5 × 10⁵ to 1 × 10⁶ each of T^CXCR4 and T^Control were mixed into a 1:1 ratio before injection, and vaccination was performed at 24 hours and on day 29 by s.c. injection of 200 μM SIINFEKL or an irrelevant peptide in a 1:1 ratio with incomplete Freund’s adjuvant (Sigma-Aldrich). Alternatively, mice received 1 × 10⁶ CD11c⁺ peptide-loaded DCs intravenously.

Khan A.B., Carpenter B., Santos e Sousa P., Pospori C., Khorshed R., Griffin J., Velica P., Zech M., Ghorashian S., Forrest C., Thomas S., Gonzalez Anton S., Ahmadi M., Holler A., Flutter B., Ramirez-Ortiz Z., Means T.K., Bennett C.L., Stauss H., Morris E., Lo Celso C, & Chakraverty R. (2018). Redirection to the bone marrow improves T cell persistence and antitumor functions. The Journal of Clinical Investigation, 128(5), 2010-2024.

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